title: Comparison of liver mitochondrial proteins derived from newborn cloned calves and from cloned adult cattle by two-dimensional differential gel electrophoresis.
abstract: Aberrant reprogramming of donor somatic cell nuclei may result in many severe problems in animal cloning. The inability to establish functional interactions between donor nucleus and recipient mitochondria is also likely responsible for such a developmental deficiency. However, detailed knowledge of protein expression during somatic cell nuclear transfer (SCNT) in cattle is lacking. In the present study, variations in mitochondrial protein levels between SCNT-derived and control cattle, and from calves derived by artificial insemination were investigated. Mitochondrial fractions were prepared from frozen liver samples and subjected to two-dimensional (2-D) fluorescence differential gel electrophoresis (DIGE) using CyDye™ dyes. Protein expression changes were confirmed with a volume ratio greater than 2.0 (P < 0.05). 2D-DIGE analysis revealed differential expression of three proteins for SCNT cattle (n = 4) and seven proteins for SCNT calves (n = 6) compared to controls (P < 0.05). Different protein patterning was observed among SCNT animals even if animals were generated from the same donor cell source. No differences were detected in two of the SCNT cattle. Moreover, there was no novel protein identified in any of the SCNT cattle or calves. In conclusion, variation in mitochondrial protein expression concentrations was observed in non-viable, neonatal SCNT calves and among SCNT individuals. This result implicates mitochondrial-related gene expression in early developmental loss of SCNT embryos. Comparative proteomic analysis represents an important tool for further studies on SCNT animals.
authors.affiliation: National Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan. kumiko@affrc.go.jp
authors.forename: Kumiko
authors.initials: K
authors.lastname: Takeda
authors.forename: Mariko
authors.initials: M
authors.lastname: Tasai
authors.forename: Satoshi
authors.initials: S
authors.lastname: Akagi
authors.forename: Shinya
authors.initials: S
authors.lastname: Watanabe
authors.forename: Mika
authors.initials: M
authors.lastname: Oe
authors.forename: Koichi
authors.initials: K
authors.lastname: Chikuni
authors.forename: Mayumi
authors.initials: M
authors.lastname: Ohnishi-Kameyama
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Hanada
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Nakamura
authors.forename: Takahiro
authors.initials: T
authors.lastname: Tagami
authors.forename: Keijiro
authors.initials: K
authors.lastname: Nirasawa
mesh_terms: D000367:Age Factors; D000818:Animals; D002417:Cattle; D002467:Cell Nucleus / Q000378:metabolism; D065150:Cellular Reprogramming; D019976:Cloning, Organism; D004624:Embryo Transfer / Q000379:methods; D005260:Female; D015870:Gene Expression; D007315:Insemination, Artificial / Q000379:methods; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D008297:Male; D008928:Mitochondria / Q000235:genetics; D024101:Mitochondrial Proteins / Q000378:metabolism; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D020543:Proteome / Q000032:analysis; D040901:Proteomics; D057825:Two-Dimensional Difference Gel Electrophoresis / Q000379:methods*
chemical_list: D024101:Mitochondrial Proteins; D020543:Proteome